Automated in situ profiling of chromatin modifications resolves cell types and gene regulatory programs

Our understanding of eukaryotic gene regulation is limited by the complexity of protein-DNA interactions that comprise the chromatin landscape and inefficient methods for characterizing these interactions. We recently introduced CUT&RUN, an antibody-targeted method that profiles DNA-binding proteins, histones and chromatin modifying proteins with high sensitivity and resolution. Here we describe an automated CUT&RUN platform and apply it to profile the chromatin landscapes of human cell lines. We develop a metric to quantitatively compare chromatin features between cell types and identify their distinctive gene expression programs. Finally, we use this method to identify gene expression features of two different pediatric diffuse midline gliomas using frozen tissue from patient-derived xenografts. Our easy, cost-effective workflow makes automated CUT&RUN an attractive tool for high-throughput characterization of cell types and patient samples. We used Cleavage under targets and Release using nuclease (Cut-and-Run), a chromatin profiling strategy in which antibody-targeted controlled cleavage by micrococcal nuclease releases specific protein-DNA complexes into the supernatant for paired-end DNA sequencing.